AgNO3 DNA Staining Procedure Questions (David Atherton, are you there?)

Mr. D.J. Atherton datherto at crc.ac.uk
Fri Jun 18 05:25:02 EST 1993


Hi Tim, Netters

My deepest and most humble apologies if I wrote to develop in Na2CO3/formamide
Developing solution should read:-

Develop in 2% Na2CO3/0.2% **FORMALDEHYDE**

I really must learn to lay off the 100% EtOH :-).

Can't think why your mail bounced except perhaps the @address I put is in 
JANET format, ie the other way around from Internet.  Failing that try just
'reply'ing when reading the mail and go ad lib.

Regards,

datherton at uk.ac.ox.path.vax
David Atherton
Univ. Oxford
Oxford
Oxon
UK
!
.

PS. Had this sent to me, thought you would like it as well.
(Sorry about cutting the .sig Ed!)

And another, from personal comm:


       RNA / DNA SILVER STAIN PROTOCOL FOR ACRYLAMIDE GELS


FROM: Moshe Balas, Volcani Centre, Rehovot, Israel

BY: Ed Rybicki, Dept Microbiology, University of Cape Town, Aug 1992

WORKS VERY WELL FOR VIROIDS, dsRNAs, AND REPUTEDLY FOR DNA ALSO


SHORT PROCEDURE:

1.   Soak in 30% EtOH / 10% acetic acid 10 min on bench, can
     leave up to O/N.

2.   Soak 10% EtOH 2x10 min, wash in ddH2O 3x5 min.

3.   Soak in 6 mM AgNO3 for 30 min (50 ml/gel), then quick rinse
     ddH2O.

4.   Rinse rapidly in fresh 2.5% Na2CO3 / 0.02% HCHO, then fresh
     soln. for 2 min or longer: develop in same solution till

     bands appear dense enough.

5.   Stop with 1% acetic and keep in same solution.

NB: CAN DESTAIN GELS WITH 2% AMMONIUM PEROXYDISULPHATE

NNB: CAN SHRINK GELS FOR STORAGE BY SOAKING IN ABSOLUTE EtOH,
     THEN ACETONE, THEN AIR-DRYING


LONG PROCEDURE:

1.   Soak polyacrylamide AFTER ELECTROPHORESIS in 50% EtOH / 10%
     acetic acid 1-3 hr or or overnight.

2.   Soak in 10% EtOH / 1% acetic acid 1 hr, rinse 3x in ddH2O.

3.   Soak in 12 mM AgNO3 1 hr, rinse 3x5 min in ddH2O.

4.   Rinse rapidly in fresh 0.75 M KOH / 0.28% HCHO, leave to
     develop in more of same solution till bands appear at
     sufficient contrast / density.

5.   Add XS H2O to stop or stop with 0.07 M Na2CO3 or 5% acetic
     acid.  Store in 1% acetic.

From:-

 | Ed Rybicki, PhD             | 
 | (ed at micro.uct.ac.za)        |



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